Expression Of ER?,GPR30 And PR In Cervical Lesions And Their Significance

?Background?Cervical cacinoma is a malignant tumor with high incidence in women over the world.Despite extensive screening and advanced treatment,there are still high rate of the morbidity and mortality.The cause of cervical cacinoma is not all clear.It is recognized that the persistent infection of Human Papillomavirus(HPV)is an important cause of cervical cacinoma.In addition,some factors,such as long-term oral contraceptives,multiple pregnancy and smoking are also the causes of cervical cacinoma.The relationship between sex hormone and HPV is unclear.Some studies have shown that there are synergistic carcinogenicity in HPV and estrogen.ER?expression is an early change in the process of cervical lesions in high-risk HPV infection.GPR30 is associated with proliferation,migration,invasion,metastasis and drug resistance of tumor cells.Few investigation suggested that the expression of GPR30 in cervical cacinoma had a good prognosis in cervical cacinoma.Progesterone can promote the carcinogenic transformation of HPV-DNA and ras-oncogenes to host cells,so as to increase the expression of E6 and E7 in tumor cells.However,the role of sex homones and the relationship with HPV infection in cervical lesions have known little,these issues remains to be further studied.In order to explore the relationship between estrogen receptor and cervical lesions and search adjuvant therapy drugs for cervical cancer.Detect Estrogen receptor family-ER?,GPR30 and progesterone receptor(PR),nuclear proliferation index(Ki-67),multiple tumor suppressor gene p16(p16)in cervical lesions,investigating the role of sex hormones in cervical lesions.To detect the effect of estrogen receptor antagonist(SERM)-Raloxifene on the growth of cancer cells.Search for adjuvant therapy or combination therapy for cervical cancer to improve the drug resistance of tumor.?Materials and methods?1.A total of 60 paraffin-embedded tissues were collected from biopsy and surgical specimens,including 10 cases of normal cervical tissue,11 cases of low grade squamous intraepithelial neoplasia(LSIL)and 21 cases of high grade squamous intraepithelial neoplasia(HSIL),squamous cell carcinoma(SCC)in 18 cases,by immunohistochemistry(IHC)staining.To detect the ER alpha,PR,GPR30,Ki67,p16 expression in different level of cervical lesions.2.Cervical squamous cell carcinoma cell lines C33-A(HPV-)and Caski(HPV16+)were selected for cell culture.Estrogen receptor antagonist(Raloxifene)was given to observe cell growth and collect cell proteins for quantitative detection of ER? expression by Western Blot,assessment of the effect of Raloxifene on the growth of cervical cancer cells.?Results?1.Immunohistochemical staining of paraffin embedded tissue:(1)ER? staining score:The normal cervical tissue average score was 10.30,LSIL average score was 9.09,HSIL average score was 6.57 and the SCC average score was3.68(P < 0.001).The difference between the four groups was statistically significant.(2)GPR30 staining score:The GPR30 score of the normal group was 2.00,LSIL average score was 3.18 and the average score of HSIL was 3.38,SCC average score was 3.63(P < 0.05).The difference between the four groups was statistically significant.(3)PR staining score:The PR average score of the normal group was 3.00,the average score of the LSIL group was 2.45,HSIL average score was 3.00,the average score of the SCC group was 2.68(P>0.05).There was no significant difference between the two groups.(4)Ki-67 staining score:The average score of normal group was 1.40,LSIL average score was 2.27 and the average score of HSIL was 3.10.The average score of SCC group was 3.47(P < 0.001).The difference between four groups was statistically significant.(5)P16 staining score:The average score of normal group was 0.001,LSILaverage score was 1.09,the average score of HSIL was 2.10 and the average score of SCC was 2.61(P<0.001).There was significant difference between the two groups.2.Results of cell drug sensitivity test:Raloxifene inhibited the expression of ER?protein in C33 A and Caski cell lines,and the cell growth was inhibited.The gray value of ER? protein in negative control group was significantly lower than that in control group(P<0.05).?Conclusions?1.The expression of ER? was related to the progression of cervical lesions.Normal cervical tissue and LSIL tissue high expression of ER?,with the gradual progress of cervical lesions ER? expression decreased,cervical cancer and high-grade lesions reduced or lacked expression of ER?;2.GPR30 was associated with the progression of cervical lesions.Normal cervical tissue and LSIL tissue was low expression or no expression,HSIL tissue and SCC was overexpression of GPR30;3.There was no significant correlation between PR expression and the degree of cervical lesion,which could not be used as an index to judge the degree of cervical lesion.4.Ki-67 and p16 were related to cervical lesions.In normal cervical tissues,P16 was not expressed and Ki-67 expressed only in basal cells of cervix.With the progression of cervical lesions,the expression of Ki-67 and P16 increased or overexpressed.5.Raloxifene inhibited the proliferation of cervical cancer cells by specifically blocking the expression of ER? in cancer cells.To sum up,the expression of ER?,GPR30,Ki-67,and p16 in cervical lesions was different.With the increase of malignant grade of cervical lesions,the expression of GPR30,Ki-67 and p16 showed an upward trend.The expression of ER? was decreased or absent with the increase of malignant degree of cervical lesions.Raloxifene can specifically reduce the expression of ER? protein and inhibit cell growth in cervical cancer cells,which is expected to be used as adjuvant therapy for cervical cancer.